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| The Effect of Leonurine on Cryopreservation of Sheep (Ovis aries) Semen |
| LI Zhi-Xing1, QING Yu-Hong1, LI Zhi-Yuan1, CAO Shao-Qi2, GAO Xin-Mei2, ZENG Wei-Bin1, WANG Jing1,*, HU Guang-Dong1,* |
1 College of Animal Science and Technology, Shihezi University, Shihezi 832003, China;
2 Animal Husbandry Station of Xinjiang Uygur Autonomous Region, Urumqi 830049, China |
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Abstract Cryopreservation of semen is one of the effective means to effectively improve the efficiency of artificial insemination in Ovis aries. Adding antioxidants to semen preservation diluent is the key to semen preservation process. Leonurine alkaloid as an antioxidant, has rarely been reported in semen preservation studies. In this study, 20, 40, 60, 80 and 100 μmol/mL leonurine alkaloid were added to the cryopreserved diluent of Duolang sheep semen, respectively. The effect of leonurine alkaloid on the cryopreservation of sheep semen was comprehensively evaluated by detecting semen quality, antioxidant capacity, relative expression of apoptotic related gene mRNA, and conception rate. The results showed that compared with the control group (0 μmol/mL), the supplementation of leonurine alkaloid in the cryoppreservation diluent could significantly increase the viability rate, acrosome integrity rate and plasma membrane integrity rate of the cryoppreservation semen of Duolang sheep (P<0.05); It significantly increased the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-PX) and catalase (CAT), increased the level of total antioxidant eapacity (T-AOC) and decreased the content of malondialdehyde (MDA)(P<0.05); The best preservation effect of leonurine alkaloid in cryopreservation was 60 μmol/mL (P<0.05); In addition, compared with the 0 μmol/mL group, the relative mRNA expressions of spermatozoa-related apoptotic genes TNF receptor superfamily, member 6 (Fas), caspase family omega-3 (Caspase-3) and BCL2-associated X (Bax) were significantly decreased (P<0.05), and the relative expression of anti-apoptotic gene B-cell lymphoma-2 (Bcl-2) was significantly increased in the optimal concentration of leonurine alkaloid group during cryopreserved (P<0.05); At the same time, the conception rate of 60 μmol/mL group was higher than that of 0 μmol/mL group, which reached the lowest level of fresh spermatozoa vaginal instillation. This study showed that adding 60 μmol /mL leonurine alkaloid to semen diluent could significantly improve the quality of frozen preserved semen of Duolang sheep, enhance its antioxidant capacity, and play an anti-apoptotic role, so that the conception rate could reach a good level. This study provides a useful reference for exploring the effect of leonurine alkaloid on sheep semen preservation and improving the effect of sheep semen preservation.
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Received: 06 October 2022
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